文档介绍：关于荧光显微镜的基本使用方法
第一页，本课件共有22页
实验目的
1. 了解荧光显微镜的基本原理；
2. 掌握荧光显微镜的基本结构和使用注意事项；
第二页，本课件共有22页
自发性荧光物质
神经细胞和心肌细胞等内的脂褐素呈棕黄色荧光。
肝贮脂细胞和视网膜色素上皮细胞内的维生素A呈绿色荧光。
某些神经内分泌细胞和神经纤维内的单***类物质（儿茶酚***、5－羟色***、组***等）在甲醛作用下呈不同颜色的荧光。
组织内含有的奎宁、四环素等药物也呈现一定的荧光。
嗜中性粒细胞中脱氢酶辅酶呈现一定的荧光。
第三页，本课件共有22页
第四页，本课件共有22页
优点：
检出能力高
对细胞的刺激小
能进行多重染色
用途：
物体构造的观察
荧光的有无、色调比较进行物质判别
发荧光量的测定对物质定性、定量分析
荧光显微镜的优点和用途
第五页，本课件共有22页
实验原理
光源
激发滤色镜
分色镜
发射滤色镜
（阻挡滤片）
第六页，本课件共有22页
实验原理
荧光显微镜的光路及主要部件
光源
激发光挡片
激发滤色镜
分色镜
发射滤色镜
第七页，本课件共有22页
第八页，本课件共有22页
Microscopes with an upright-style frame are capable of producing fluorescence illumination either through episcopic or diascopic optical pathways, although the latter is rarely used today. Epi-illuminators usually consist of a mercury or xenon lamphouse coupled to a vertical illuminator that is positioned above the main frame in a separate assembly. The microscope nosepiece and transmitted light components (diascopic illuminator, condenser, field diaphragm, filters, etc.) are built into the main frame, while fluorescence components are housed in the vertical illuminator. These illuminators often contain a revolving or sliding turret that houses four to six "cubes" that contain a mixture of interference filters including a barrier filter, dichroic mirror, and an excitation filter. As illustrated above, light emitted from the lamp positioned in the episcopic lamphouse passes through a collector lens and then the field and aperture diaphragms before entering the first interference filter in the cube set, the emission filter. This light is then directed through the objective and onto the specimen by a special dichroic mirror that reflects certain wavelengths while passing others. Secondary fluorescence, emitted by fluorophores residing in the specimen, travels back through the objective and the dichroic mirror before passing through a barrier filter and into the microscope eyepieces or camera system.
第九页，本课件共有22页
激发光源
高压***灯拥有313, 337, 365, 405, 436, 5